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Circular permutation of red fluorescent proteins.

ABSTRACT: Circular permutation of fluorescent proteins provides a substrate for the design of molecular sensors. Here we describe a systematic exploration of permutation sites for mCherry and mKate using a tandem fusion template approach. Circular permutants retaining more than 60% (mCherry) and 90% (mKate) brightness of the parent molecules are reported, as well as a quantitative evaluation of the fluorescence from neighboring mutations. Truncations of circular permutants indicated essential N- and C-terminal segments and substantial flexibility in the use of these molecules. Structural evaluation of two cp-mKate variants indicated no major conformational changes from the previously reported wild-type structure, and cis conformation of the chromophores. Four cp-mKates were identified with over 80% of native fluorescence, providing important new building blocks for sensor and complementation experiments.

SUBMITTER: Shui B

PROVIDER: S-EPMC3103546 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Circular permutation of red fluorescent proteins.

Shui Bo B Wang Qi Q Lee Frank F Byrnes Laura J LJ Chudakov Dmitry M DM Lukyanov Sergey A SA Sondermann Holger H Kotlikoff Michael I MI

PloS one 20110527 5

Circular permutation of fluorescent proteins provides a substrate for the design of molecular sensors. Here we describe a systematic exploration of permutation sites for mCherry and mKate using a tandem fusion template approach. Circular permutants retaining more than 60% (mCherry) and 90% (mKate) brightness of the parent molecules are reported, as well as a quantitative evaluation of the fluorescence from neighboring mutations. Truncations of circular permutants indicated essential N- and C-ter ...[more]

PMID: 21647365

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